Specific Noncompetitive Immunoassay for HT-2 Mycotoxin Detection

Analytical Chemistry
Henri O ArolaTarja K Nevanen

Abstract

Here we demonstrate a novel homogeneous one-step immunoassay, utilizing a pair of recombinant antibody antigen-binding fragments (Fab), that is specific for HT-2 toxin and has a positive readout. Advantages over the conventional competitive immunoassay formats such as enzyme-linked immunosorbent assay (ELISA) are the specificity, speed, and simplicity of the assay. Recombinant antibody HT2-10 Fab recognizing both HT-2 and T-2 toxins was developed from a phage display antibody library containing 6 × 10(7) different antibody clones. Specificity of the immunoassay was introduced by an anti-immune complex (IC) antibody binding the primary antibody-HT-2 toxin complex. When the noncompetitive immune complex assay was compared to the traditional competitive assay, an over 10-fold improvement in sensitivity was observed. Although the HT2-10 antibody has 100% cross-reactivity for HT-2 and T-2 toxins, the immune complex assay is highly specific for HT-2 alone. The assay performance with real samples was evaluated using naturally contaminated wheat reference material. The half-maximal effective concentration (EC50) value of the time-resolved fluorescence resonance energy transfer (TR-FRET) assay was 9.6 ng/mL, and the limit of detection (...Continue Reading

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Citations

Jul 13, 2016·International Journal of Biological Macromolecules·Rasmus P ClausenPeter D Skottrup
Apr 21, 2017·Toxins·Henri O ArolaTarja K Nevanen
Nov 12, 2017·Analytical and Bioanalytical Chemistry·Riikka PeltomaaMaría C Moreno-Bondi
Jan 1, 2020·Scientific Reports·F WuA N Grigorenko
Jul 27, 2021·Frontiers in Cellular and Infection Microbiology·Kristian Daniel Ralph RothMichael Hust
Feb 23, 2019·Analytical Chemistry·Jinjie LiHeyou Han
Aug 29, 2019·ACS Omega·Riikka PeltomaaMaría C Moreno-Bondi
Aug 22, 2021·Analytical and Bioanalytical Chemistry·Riikka PeltomaaMaría C Moreno-Bondi
Jan 4, 2022·Analytical and Bioanalytical Chemistry·Fernando Pradanas-GonzálezMaría C Moreno-Bondi

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